Caenorhabdtis elegans is an important model organism for identifying and studying druggable targets in a high-throughput manner [1–3]. We have previously described a modular pipeline for image processing and analysis for both parasitic and free-living worms . Here, we detail the upstream assay set up for high throughput drug screens of the free-living nematode C. elegans.
Following a standard bleaching protocol, embryos are hatched overnight and synchronized L1s are added to a 96-well plate. Concentrated Esherichia coli strain HB101 bacterial food  is added to each well to sustain growth for 48 hours along with compounds of interest. Plates are incubated for 48 hours until worms reach the young adult stage. While this assay has been optimized for image acquisition with an ImageXpress high-content imager, it can be adapted for use on similar imaging platforms. Analysis of images from an ImageXpress is seamless using wrmXpress , but other analysis software can be used.